Psychedelic tryptamines such as DMT, 5-MeO-DMT, and bufotenine, found naturally in plants and animals, show clinical promise for treating anxiety and depression. Using genetic and metabolic engineering, researchers developed a biosynthetic pathway in Escherichia coli to produce these compounds. With tryptophan supplementation, DMT reached maximum titers of 74.7 ± 10.5 mg/L in fed-batch 2-L bioreactors. De novo DMT production from glucose achieved 14.0 mg/L, and the study reports the first microbial production of 5-MeO-DMT and bufotenine in vivo. This work establishes a foundation for further optimization toward industrial-scale production.
N-methylated tryptamines like psilocybin and DMT show promise as treatments for mental health disorders, driving interest in biosynthetic production. This work characterized two enzymes from tryptamine biosynthesis: TrpM, a tryptophan N-methyltransferase from Psilocybe serbica, and PsiD, a decarboxylase from the psilocybin pathway. TrpM was able to N-methylate 4-hydroxytryptophan, a non-native amino acid. However, incorporating TrpM into a functional psilocybin pathway was blocked because PsiD could not use N,N-dimethyl-4-hydroxytryptophan as a substrate under the tested conditions, despite acting on N-methylated and 4-hydroxylated tryptophan derivatives separately. These findings expand the known substrates for TrpM and PsiD, increasing the diversity of tryptamine biosynthetic products.